卵巢癌组织MEOX1表达对细胞增殖和侵袭及迁移影响

目的卵巢癌极易发生转移和耐药,严重影响患者预后。前期研究发现,同源盒基因MEOX1可能在肿瘤的转移中发挥着重要作用。但有关MEOX1在卵巢癌中的表达和功能尚罕见报道。本研究探讨MEOX1对卵巢癌细胞增殖、迁移和侵袭的影响,及其在卵巢癌组织中的表达水平和临床意义。方法采用脂质体转染法将靶向MEOX1的特异性siRNA序列、无效序列分别转染至人卵巢癌细胞OAW28中,命名为实验(siMEOX1)组和阴性对照(siNC)组,同时设置空白对照(Control)组。采用实时荧光定量PCR(real-time fluorescence quantitative PCR,qPCR)和蛋白质印迹法检测转染siRNA对人卵巢癌细胞OAW28中MEOX1的敲降效果。采用Transwell小室法检测细胞的体外迁移和侵袭能力。采用CCK8法检测细胞的体外增殖能力。采用免疫组织化学染色法检测购于西安艾丽娜生物科技有限公司的73例卵巢癌组织中MEOX1的表达水平。结果qPCR和蛋白质印迹结果显示,siMEOX1转染后,卵巢癌OAW28细胞中MEOX1表达水平明显降低,RNA水平敲降效率约72%。敲降MEOX1后卵巢癌OAW28细胞的增殖能力受到抑制,差异有统计学意义,F=59.217,P<0.001。此外迁移实验显示,siMEOX1组迁移细胞数(108.80±4.27)少于siNC组(230.20±5.63),差异有统计学意义,F=491.571,P<0.001。侵袭实验显示,siMEOX1组侵袭细胞数(217.80±3.49)低于siNC组(379.80±3.96),差异有统计学意义,F=712.267,P<0.001。免疫组化结果显示,MEOX1在卵巢癌组织中表达阳性率为60.3%(44/73)。临床关联性研究发现,MEOX1阳性表达与卵巢癌患者的转移具有关联性,χ^2=7.637,P=0.004;但与患者的年龄、TNM分期、病理分级无关联。结论MEOX1可促进人卵巢癌细胞的体外增殖和迁移侵袭能力,且MEOX1高表达与卵巢癌患者转移具有关联性,可能成为抑制卵巢癌转移的潜在靶点。

Expression of MEOX1 in ovarian cancer and its effect on the proliferation,invasion and migration

OBJECTIVE The mortality rate of ovarian cancer is the first in the gynecological tumor.Ovarian cancer is highly susceptible to metastasis and drug resistance,which seriously affects the patient’s prognosis.The previous studies indicated that the homeobox gene MEOX1 could play an important role in the metastasis of the tumor.However,the expression and function of MEOX1 in ovarian cancer have not been reported.In this study,we not only studied the effect of MEOX1 on human ovarian cancer cell,but also investigated the its expression and clinicopathological relevance in human ovarian cancer tissues.METHODS Specific siRNA sequence targeting MEOX1 and null sequence were transfected into human ovarian cancer cell line OAW28 by lipofection,named as experimental(siMEOX1)group and negative control(siNC)group,and the blank control(control)group was simultaneously set.Real-time fluorescence quantitative PCR(qPCR)and Western blot were used to detect the levels of MEOX1 in OAW28 cells after siRNA transfected.Transwell assay was used to detect the migration and invasion of cells in vitro.And CCK8 assay was used to measure cell proliferation in vitro.The expression of MEOX1 in 73 cases of ovarian cancer tissues was detected by immunohistochemisty using the corresponding primary antibody on one commercial tissue array.RESULTS qPCR and Western blot demonstrated that siMEOX1 significantly decreased the expression of MEOX1 in OAW28 cells,the knockdown efficiency of RNA level was about 72%.Silencing of MEOX1 by specific siRNA significantly inhibited OAW28 cell proliferation(F=59.217,P<0.001).In Transwell migration assay,the number of migrated cells of siMEOX1 group was 108.80±4.27 per sight,while the Negative Control group was 230.20±5.63(F=491.571,P<0.001).Similarly,the invasiveness capability of cells in the siMEOX1 group(217.80±3.49)was significantly decreased as compared with that in Negative Control group(379.80±3.96)in Transwell invasion assay(F=712.267,P<0.001).Immunohistochemical results showed that the positive rate of MEOX1 in ovarian cancer tissues was 60.3%(44/73).The Clinicopathological relevance analysis showed that the high levels of MEOX1 was significantly associated with metastasis(χ^2=7.637,P=0.004),but there was no significant difference with the age,TNM stage and pathological grade.CONCLUSIONS MEOX1 could promote the proliferation,migration and invasion of human ovarian cancer cell OAW28 in vitro.The positive expression of MEOX1 in ovarian cancer tissues is closely associated with metastasis,which could be the candidate target for inhibiting the metastasis of ovarian cancer.